The expressions of AIM-1 and STK15 during the developmental processes of normal megakaryocytes. CD34+ cells were purified from normal human bone marrow cells, and then cultured with TPO (20 ng/ml) and PPP (20%) for the indicated times. (A) Phase–contrast micrographs of cells before and after the culture. Bar, 20 μm. (B) Light micrographs of cells. Cytocentrifugation preparations of the cells before and after the culture were stained with May-Grunwald-Giemsa. Bar, 10 μm. (C) DNA content of cultured cells was analyzed by flow cytometry before (dashed line) and after (solid line) culture. (D) Semiquantitative RT-PCR analysis on the expression of AIM-1 and STK15 mRNA. Total cellular RNA was isolated at the times indicated and cDNA was synthesized. The amounts of cDNA products were normalized according to the amounts of PCR products of GAPDH. The adjusted amounts of cDNA products from each sample were subjected to the PCR reaction for AIM-1, STK15, GPIIb, and GAPDH. The PCR products were size fractionated on 3.5% polyacrylamide gels, dried, and autoradiographed. NTC, no template control.
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